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Cell Line Development Using Targeted Gene Integration into MAR-Rich Landing Pads for Stable Expression of Transgenes

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Genotype Phenotype Coupling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2681))

Abstract

Integration of a gene of interest (GOI) into the genome of mammalian cells is the first step of cell line development campaigns for the production of biotherapeutics. Besides random integration methods, targeted gene integration approaches have emerged as promising tools over the last few years. In addition to reducing heterogeneity within a pool of recombinant transfectants, this process can also facilitate shorter timelines of the current cell line development process. Herein, we describe protocols for generating host cell lines carrying matrix attachment region (MAR)-rich landing pads (LPs), including BxB1 recombination sites. These LP-containing cell lines allow for site-specific and simultaneous integration of multiple GOIs. The resulting transgene-expressing stable recombinant clones can be used for the production of mono- or multispecific antibodies.

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Author information

Authors and Affiliations

  1. Institute of Life Technologies, Haute Ecole d’Ingénierie HES-SO Valais Wallis, Sion, Switzerland

    Claudia Oliviero, Steffen C. Hinz & Gerrit Hagens

  2. Ferring Biologics Innovation Centre, Epalinges, Switzerland

    Julius Grzeschik

  3. Aerium Therapeutics, Epalinges, Switzerland

    Björn Hock

  4. Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany

    Harald Kolmar

Authors
  1. Claudia Oliviero
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  2. Steffen C. Hinz
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  3. Julius Grzeschik
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  4. Björn Hock
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  5. Harald Kolmar
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  6. Gerrit Hagens
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Corresponding author

Correspondence to Gerrit Hagens .

Editor information

Editors and Affiliations

  1. Protein Engineering and Antibody Technologies (PEAT), Merck Healthcare KGaA, Darmstadt, Germany

    Stefan Zielonka

  2. Protein Engineering and Antibody Technologies (PEAT), Merck Healthcare KGaA, Darmstadt, Germany

    Simon Krah

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Oliviero, C., Hinz, S.C., Grzeschik, J., Hock, B., Kolmar, H., Hagens, G. (2023). Cell Line Development Using Targeted Gene Integration into MAR-Rich Landing Pads for Stable Expression of Transgenes. In: Zielonka, S., Krah, S. (eds) Genotype Phenotype Coupling. Methods in Molecular Biology, vol 2681. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3279-6_19

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  • DOI: https://doi.org/10.1007/978-1-0716-3279-6_19

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3278-9

  • Online ISBN: 978-1-0716-3279-6

  • eBook Packages: Springer Protocols

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